This research project builds on existing transgenic and cloning expertise using nonhuman primates (NHP) and NIH-approved human embryonic stem cells (hES cells) to develop novel Alzheimer disease models. We propose generate transgenic AD-models in hES cells, as well as non-human primate embryonic stem cells (nhpES cells), using lentiviral vectors (LV: Aim I) and mammalian artificial chromosomes (MAC: Aim II). The ES cells let us perform differentiation into AD neural tissues in vitro by determining amyloid overexpression in h/nhpES differentiated into neuroprogenitor cells (NP) in vitro, in three-dimensional embryoid body cultures (EB) and in xenografts into SCID mice. The cell lines, embryoid bodies and xenografts generated will be characterized for amyloid overexpression, plaque formation and neurofibrillary tangles. Aim III, investigated exclusively in nonhuman primates in accordance with US Federal guidelines, marries AD-transgenesis with cloning using both somatic cell nuclear transfer (SCNT) or embryonic stem cell nuclear transfer (ESNT) for the purpose of deriving nonhuman primate embryonic stem cells (NT-nhpES cells), to be tested in vitro, in xenografts and after cell transplantation to determine whether undifferentiated NT-nhpES or differentiated NT-nhpES neuroprogenitors are tolerated by the host monkey. By year five these experiments will provide proof of principle that transgenic AD-primate embryos can be generated through NT eventually leading to the live births of NHP-AD offspring. Overall, this interdisciplinary research project develops novel in vitro and the proof of principle for preclinical AD models.